hanged daily, and spontaneously differentiated colonies were removed when appropriate. iPS cells were passaged every six days with 0.05% trypsin-EDTA at 37uC for 35 min. When colonies near the edge of the plate began to dissociate from the bottom, the enzyme was removed, and colonies were washed with mTeSR1 medium. Cells were collected by gently pipetting and replated onto fresh Matrigel-coated dishes. The ROCK inhibitor Y-27632 was added to each well for the first day after each passage. Annexin V and propidium iodide assays iPS cells were cultured in mTeSR1 medium in Piceatannol cost 6-well plates to produce colonies at 80%90% confluence. The iPS cells were irradiated with UVC as described above. The UVC+30 nM maxadilan iPS samples were treated with 30 nM of maxadilan for 1 h prior to exposure to 100 J/m2 UVC, and the UVC+0 nM maxadilan iPS samples were exposed to 100 J/m2 UVC in the absence of maxadilan. After iPS cells were exposed 10595516” to UVC, fresh culture medium and the appropriate concentration of maxadilan were added to each well, and the cells were incubated for 7 h. Control wells containing iPS cells were cultured in mTeSR1 medium without UVC irradiation. To detect early apoptotic activity, an Annexin V-FITC/PI Apoptosis Detection Kit was used according to the manufacturer’s instructions. iPS cells were washed with cold PBS and added to 200 ml of the Annexin V-binding buffer. After the samples were stained with 2 ml of FITC-labeled Annexin V and 2 ml of PI, the samples were immediately analyzed by flow cytometry. Western blot analysis PAC1 was detected by western blot analysis in iPS cells. iPS cells were pretreated with 100 nM of maxadilan for 24 h and passaged 3 times without removing the spontaneously differentiated colonies prior to quantitative western blot analysis for Nanog, OCT4 and 11948668” SOX2 protein levels. This same procedure was used on control iPS cells that were not treated with maxadilan. iPS cells were lysed with RIPA buffer containing a protease inhibitor cocktail and sonicated on ice. The sonicated material was then centrifuged for 20 min at 15,0006rpm at 4uC, and the supernatant was collected. Fifty Micrograms of total protein as determined by the BCA method were separated on 10% SDS-PAGE gels and transferred onto a PVDF membrane. The membrane was incubated overnight at 4uC with the following primary antibodies: rabbit polyclonal anti-PAC1 antibody, rabbit polyclonal anti-OCT4 antibody, rabbit polyclonal anti-Nanog antibody, rabbit monoclonal anti-SOX2 antibody and rabbit monoclonal anti-b-actin antibody. The membrane was incubated with the goat anti-rabbit IgG secondary antibody for 1 h 
at room temperature. After adding the ECL chemiluminescence reagent, the membrane was incubated with developer solution for 1 min and with a fixative for 0.5 min. Quantitation of band Terminal transferase dUTP nick end labeling assays iPS cells were cultured in mTeSR1 medium in 6-well plates to produce colonies at 80%90% confluence and irradiated with UVC as described above. The UVC+30 nM maxadilan iPS sample was treated with 30 nM maxadilan for 1 h prior to exposure to 100 J/m2 UVC, whereas the UVC+0 nM maxadilan iPS group was exposed to 100 J/m2 UVC without pretreatment with maxadilan. After the iPS cells were exposed to UVC, fresh culture medium and the appropriate concentration of maxadilan were added to each well, and the cells were incubated for 9 h. Control wells contained iPS cells cultured in mTeSR1 medium and did not receive UVC radiation. T